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MOLECULAR AND CELLULAR BIOLOGY, June 2007, p. 3995–4005
Copyright © 2007, American Society for Microbiology. All Rights Reserved.
Vol. 27, No. 11
A Double-Stranded-RNA Response Program Important for
RNA Interference Efficiency?
Swati Choudhary,1† Heng-Chi Lee,1† Mekhala Maiti,1† Qun He,1‡ Ping Cheng,1§
Qinghua Liu,2and Yi Liu1*
Department of Physiology1and Department of Biochemistry,2University of Texas Southwestern Medical Center,
5323 Harry Hines Blvd., Dallas, Texas 75390
Received 31 January 2007/Returned for modification 6 March 2007/Accepted 13 March 2007
When recognized by the RNA interference (RNAi) pathway, double-stranded RNA (dsRNA) produced in
eukaryotic cells results in posttranscriptional gene silencing. In addition, dsRNA can trigger the interferon
response as part of the immune response in vertebrates. In this study, we show that dsRNA, but not short
interfering RNA (siRNA), induces the expression of qde-2 (an Argonaute gene) and dcl-2 (a Dicer gene), two
central components of the RNAi pathway in the filamentous fungus Neurospora crassa. The induction of QDE-2
by dsRNA is required for normal gene silencing, indicating that this is a regulatory mechanism that allows the
optimal function of the RNAi pathway. In addition, we demonstrate that Dicer proteins (DCLs) regulate QDE-2
posttranscriptionally, suggesting a role for DCLs or siRNA in QDE-2 accumulation. Finally, a genome-wide
search revealed that additional RNAi components and homologs of antiviral and interferon-stimulated genes
are also dsRNA-activated genes in Neurospora. Together, our results suggest that the activation of the RNAi
components is part of a broad ancient host defense response against viral and transposon infections.
The production of double-stranded RNA (dsRNA) in eu-
karyotic cells, generally as the result of viral replication or the
transcription of transposable elements and repetitive DNA
sequences, is known to elicit two types of cellular defense
responses. In the first type of response, dsRNA is recognized
and cleaved by the RNase III enzyme Dicer to yield 20- to
25-nucleotide short interfering RNA (siRNA) duplexes (5, 38).
The production of siRNA initiates RNA interference (RNAi),
also known as posttranscriptional gene silencing. The siRNAs
are loaded onto the RNA-induced silencing complex (RISC),
of which an Argonaute (Ago) family protein forms the catalytic
core, and guide the RISC to recognize and cleave homologous
mRNA or result in the transcriptional silencing of the homol-
ogous DNA locus (31, 43–45, 49, 53). Dicer and the Argonaute
proteins are the central components of the RNAi pathway.
The RNAi pathway plays important roles in silencing trans-
poson and viruses in animal and plant cells and is thought to be
a defense system against viruses and transposons (20, 30, 33,
46, 56). Components of the RNAi pathway probably cleave
viral dsRNA and degrade viral and transposon mRNAs. Con-
sistent with this notion, many viral genomes are known to
encode inhibitors of the RNAi pathway (6, 20, 34).
In vertebrates, dsRNA and siRNA are also known to trig-
ger the transcription-based antiviral interferon (IFN) response
(24, 27, 47). In mammalian cells, dsRNA is recognized by
dsRNA sensors such as Toll-like receptor 3 and the dsRNA-
dependent protein kinase R (PKR). This leads to the activa-
tion of the IFN-regulatory transcription factors and NF-?B,
which in turn results in the expression of the IFNs. The ex-
pression of IFNs then activates the transcription of hundreds
of IFN-stimulated genes (ISGs) through the JAK-STAT path-
way (19). Many of the ISGs encode proteins with antiviral
activities, including PKR and myxovirus (influenza virus) re-
sistance (Mx) proteins (23, 24, 40, 41). Thus, the IFN response
plays a crucial role in antiviral immunity in vertebrates.
Although the basic mechanism of RNAi is now fairly well
understood, little is known regarding the regulation of the
RNAi components. It is also not known whether there is cross
talk between the transcription-based antiviral response and the
posttranscription-based RNAi pathway. In addition, although
the dsRNA-induced IFN response has been well characterized,
it has been documented only in vertebrate systems, and the
evolutionary root of the response is not known.
The filamentous fungus Neurospora crassa was one of the
first model systems used for RNAi studies (15, 16); in Neuro-
spora, the RNAi pathway is essential for gene silencing (also
called quelling) induced by dsRNA or transgenes (9). Genetic
analysis led to the identification of three quelling-deficient
genes (qde's): qde-1, qde-2, and qde-3. QDE-1 (an RNA-de-
pendent RNA polymerase) and QDE-3 (a DNA helicase) are
thought to function upstream of the RNAi pathway and are
involved in the generation of dsRNA from aberrant RNAs (15,
17). Consistent with this notion, QDE-1 and QDE-3 are not
required for the production of siRNA and gene silencing if
dsRNA is made from an exogenous hairpin RNA (10). QDE-2
is an Argonaute protein and is the core of the RISC complex
associated with siRNA (7, 8, 35). In addition, Neurospora has
two partially redundant Dicer proteins, DCL-1 and DCL-2;
DCL-2 is responsible for most of the siRNA-generating activ-
* Corresponding author. Mailing address: Department of Physiol-
ogy, Room ND13.214A, University of Texas Southwestern Medical
Center, 5323 Harry Hines Blvd., Dallas, TX 75390-9040. Phone: (214)
645-6033. Fax: (214) 645-6049. E-mail: email@example.com.
† S.C., H.-C.L., and M.M. contributed equally to this study.
‡ Present address: State Key Laboratory for Agro-Biotechnology,
College of Biological Sciences, Chinese Agricultural University, Bei-
jing 100094, People's Republic of China.
§ Present address: Merck Research Laboratories, 126 E. Lincoln
Ave., Rahway, NJ 07065.
?Published ahead of print on 19 March 2007.
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ity (10). The disruption of both dcl genes in Neurospora abol-
ished all siRNA production in vivo and in vitro. As in animals,
the RNAi pathway has been shown to be important for trans-
poson silencing in Neurospora (39).
To understand the regulation of the RNAi pathway, we
examined the responses of the RNAi components after the
induction of dsRNA. dsRNA induced the transcription of both
qde-2 and dcl-2, two of the central components of the RNAi
pathway. In addition, the accumulation of the QDE-2 protein
after the induction of dsRNA required the presence of the
DCLs, suggesting that either the DCLs or the produced siRNA
regulates the steady-state level of QDE-2 posttranscriptionally.
Importantly, we showed that the induction of QDE-2 by
dsRNA is required for efficient RNAi. Finally, using microar-
ray and quantitative PCR (qPCR) analyses, we identified 60
dsRNA-activated genes (DRAGs) in Neurospora. Functional
classification of the DRAGs suggests that the induction of
RNAi components is a part of a broad host defense response
against viral infection and transposons in this filamentous
Strains and growth conditions. The wild-type strain used in this study was
either FGSC987 (A; obtained from Fungal Genetic Stock Center) or 87-3 (bd a).
Either FGSC462 (his-3 A) or 301-6 (bd his-3 A) was the host strain used for the
insertion of the his-3-targeting constructs. The following mutant strains (in the
wild-type or his-3 background) were created for this study as described below:
dcl-1ko, dcl-2RIP, dcl-1kodcl-2RIP, qde1ko, qde2RIPand qde3ko. The mutants in the
his-3 background were used as the host strains for his-3-targeting constructs.
Culture conditions were the same as those described previously (1), by inocu-
lating mycelium mats in shaking flasks with 50 ml medium. For liquid cultures
containing quinic acid (QA), 0.01 M QA (pH 5.8) was added to the liquid culture
medium containing 1? Vogel's medium, 0.1% glucose, and 0.17% arginine (13).
The cultures were grown in the presence of QA for 2 days unless otherwise
indicated. For liquid cultures containing histidine, a final concentration of 0.5
mg/ml was used.
Creation of mutant strains. The Neurospora qde-2 and dcl-2 genes were dis-
rupted by a repeat-induced point mutation (RIP) (4). The PCR fragment con-
taining the entire qde-2 or dcl-2 open reading frame (ORF) and its 3? untrans-
lated region (3.3 kb for qde-2 and 5.1 kb for dcl-2) was cloned into pDE3BH and
introduced into the his-3 locus of a wild-type strain (7088 his-3 a) by electropo-
ration (36). A positive transformant was crossed with a wild-type strain. DNA
sequencing was performed to identify the strains in which the endogenous qde-
2/dcl-2 ORF was mutated with multiple premature stop codons.
The gene replacement method was used to disrupt the Neurospora qde-1,
qde-3, and dcl-1 genes. A PCR fragment containing the entire ORF and 3?
untranslated region of the gene (qde-1/qde-3/dcl-1) was cloned into pDE3BH,
resulting in pQDE-1/pQDE-3/pDCL-1.
To make the disruption construct, a hygromycin resistance gene (hph) frag-
ment containing promoter and terminator sequences was inserted into the XbaI-
PvuII site of pQDE-1, the PvuII site of pQDE-3, or the BamHI site of pDCL-1.
A PCR fragment containing the gene replacement cassette was introduced into
a wild-type strain by electroporation (36) to select for hygromycin-resistant
transformants (200 ?g/ml hygromycin). PCR was performed to identify strains
carrying the hph fragment at the endogenous locus. Positive transformants were
crossed with a wild-type strain, and sexual spores were picked individually and
germinated on slants containing hygromycin. Southern blot analysis was per-
formed to confirm the qde-1ko, qde-3ko, and dcl-1kostrains. A dcl-1kostrain was
crossed with a dcl-2ripstrain to generate a dcl-1kodcl-2ripdouble mutant. All
mutants were crossed with a his-3 strain to obtain mutants in the his-3 back-
ground; these mutants were used for all his-3-targeting transformations.
Creation of dsRNA strains. dsRNA constructs were created as described
previously (11). The following regions from al-1, frq, and frh were cloned into
pDE3BH.qa in reverse and forward orientations, respectively: al-1, bp 1322 to
1942 and bp 1412 to 1942; frq, bp 669 to 2309 and bp 791 to 1252; and frh, bp 2087
to 2703 and bp 2189 to 2703. The first nucleotide of each ORF is counted as 1.
The resulting plasmids that contain the inverted repeats under the control of the
qa-2 promoter were targeted to the his-3 locus of a wild-type strain (301-6 bd
his-3 A) and other his-3 RNAi mutant strains by transformation (36). The
construct that contains the wild-type qde-2 gene with its own promoter was
created as described previously (35). To create plasmids used for qde-2 promoter
analyses, PCR fragments of qde-2 were inserted into the NdeI site of the double-
stranded al-1 (dsal-1) construct. Primer sequences are available upon request.
Antibody generation. A glutathione S-transferase–QDE-2 (containing QDE-2
amino acids 541 to 938) fusion protein was expressed in BL21 cells, and inclusion
bodies containing the recombinant proteins were purified and used as the anti-
gen to generate rabbit polyclonal antiserum as described previously (12).
Dicer-2-specific antibody was generated by using a DCL-2-specific peptide
(DRDDSSQDPDDNESF) synthesized by the peptide synthesis facility at the
University of Texas Southwestern Medical System. The polyclonal antiserum
from rabbit was purified using a DCL-2 peptide-conjugated affinity column.
Quelling assay. The quelling assay was performed in a wild-type strain by
cotransforming a 1.5-kb PCR fragment of the al-1 ORF and pBT6 (a benomyl-
resistant gene-containing plasmid [obtained from the Fungal Genetic Stock
Center {FGSC}]). Benomyl-resistant transformants were picked to identify yel-
low (partially quelled) or white (fully quelled) strains. The two primers used to
generate the al-1 PCR fragment were al-1-1for (5?-CTTCCGCCGCTACCTCT
CGTGG-3?) and al-1-2 rev (5?-CCCTTTGTTGGTGGCGTTGATG-3?). The
experiments were performed in constant white luminescence light at room tem-
Protein and RNA analyses. Protein extraction, quantification, and Western
blot analysis were performed as previously described (12). Equal amounts of
total protein (50 ?g) were loaded into each protein lane, and after electrophore-
sis, proteins were transferred onto a polyvinylidene difluoride membrane, and
Western blot analysis was performed. An amido black-stained membrane or
nonspecific cross-reacting bands were used as loading controls.
RNA extraction and Northern blot analyses were performed as described
previously (1). Equal amounts of total RNA (30 ?g) were loaded onto agarose
gels for electrophoresis, and the gels were blotted and probed with an RNA
probe specific for qde-2 or dcl-2. The rRNA levels were shown by the ethidium
bromide-stained agarose gels.
Microarray analysis. Neurospora oligonucleotide microarray chips were ob-
tained from the FGSC. The microarray (created by the Neurospora Genome
Project) consists of 10,526 predicted ORFs, which covers nearly all predicted
genes in Neurospora. The chips were postprocessed and UV cross-linked as
suggested by the manufacturer (GAPS II coated slides; Corning). Microarray
experiments, including aminoallyl cDNA synthesis, CyDye conjugation, and ar-
ray hybridization, were performed using Pronto Plus Indirect systems (Promega).
Cy5 and Cy3 (CyDye; Amersham Biosciences) were used to label experimental
and control cDNAs, respectively. Microarray spot analysis and acquisition were
performed using a GenePix4000B scanner with GenePix6 software (Axon In-
struments). GeneTraffic software (Stratagene) was used for normalization and
further data analysis.
qRT-PCR. Quantitative real-time PCR (qRT-PCR) was performed with an
Applied Biosystems Prism 7900HT sequence detection system using a previously
described protocol (29). Briefly, total RNA was purified using an RNeasy mini kit
(QIAGEN) and treated with DNase I (0.6 units; Roche). Equal amounts of
DNase-treated RNAs (2 ?g) were reverse transcribed with SuperScript II (In-
vitrogen) using random hexamers. cDNAs (50 ng) were mixed in 10 ?l qRT-PCR
mix with 5 ?l SYBR Green PCR Master Mix (Applied Biosystems) and 150 nM
primers. Each reaction was duplicated, and non-reverse-transcribed samples
were used as controls. Gene-specific primers were designed using Primer Express
software (Perkin-Elmer Life Sciences), and each primer pair was validated by
cDNA template titration to ensure similar amplification kinetics and a single
melting point of quantitative PCR products. Primer sequences are available upon
request. Levels of the housekeeping gene beta-tubulin were used to calculate
changes (n-fold) by comparing mean threshold cycle values.
Induction of qde-2 expression by dsRNA. Since QDE-2, the
Argonaute-like protein, is a core component of the Neurospora
RNAi pathway, we hypothesized that its expression should be
highly regulated. To test this possibility, we examined QDE-2
protein levels using a QDE-2-specific antibody in Neurospora
RNAi mutants in which one or more of the RNAi genes were
disrupted. As shown in Fig. 1A, levels of QDE-2 were reduced
in the dcl-1kodcl-2ripdouble mutant and in the qde-1koand
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qde-3kosingle mutants compared to those in the wild-type
strain. No QDE-2 protein was detected in the qde-2ripstrain,
indicating the specificity of our QDE-2 antibody. These data
indicate that the expression of QDE-2 is regulated by other
components of the RNAi pathway. Since QDE-1 and QDE-3
are involved in the synthesis of endogenous dsRNA from ab-
errant RNAs, and DCLs are responsible for the production of
siRNA, this suggests that the production of dsRNA or siRNA
promotes the expression of QDE-2 proteins.
We then examined the QDE-2 levels in albino-1 (al-1, in-
volved in carotenoid biosynthesis)-quelled strains. In these
strains, al-1 was silenced by the introduction of multiple copies
of an al-1 DNA fragment, resulting in yellow (partially quelled)
or white (fully quelled) conidia and hyphae (data not shown).
Since quelling involves the production of dsRNA, the degree
of al-1 silencing may reflect the amounts of dsRNA produced.
As shown in Fig. 1B, compared with levels observed in the
wild-type strain (orange conidia), QDE-2 levels were increased
in two partially quelled strains and further increased in two
fully quelled strains, suggesting that the production of dsRNA
in these strains leads to the increase in QDE-2 expression.
To directly investigate the effect of dsRNA on QDE-2, we
examined whether the induction of dsRNA alone was sufficient
for the increase in QDE-2 expression. Previously, we and oth-
ers developed a method to inducibly express dsRNA from
inverted repeat sequences to silence gene expression in Neu-
rospora (11, 22). The expression of the inverted repeats of the
gene of interest are controlled by the QA-inducible (qa-2)
promoter (21), so that the addition of QA to the medium will
lead to the production of dsRNA and initiate gene silencing.
The expression of dsRNA leads to the production of homolo-
gous siRNA in the wild-type strain but not in the dcl double
mutant (10, 35). QDE-2 levels were examined in wild-type and
qde-2ripstrains and in three wild-type transgenic strains, dsal-1,
dsfrq, and dsfrh, which carry constructs to induce dsRNA spe-
cific for al-1, frequency (frq, a circadian clock gene), and frh (an
essential RNA helicase gene required for circadian clock func-
tion), respectively. As shown in Fig. 1C, the presence of QA led
to a significant increase in QDE-2 levels in all three strains
with the dsRNA constructs. In contrast, QA had no effect on
QDE-2 in the wild-type strain. Since these strains produce
different dsRNAs, the effect on QDE-2 is dependent on the
production of dsRNA rather than gene-specific dsRNA. In
addition, we found that the QDE-2 levels in strains with a
mutated al-1 gene were comparable to that of the wild-type
strain, indicating that carotenoid biosynthesis is not involved in
the regulation of QDE-2 expression (data not shown).
To determine whether the increase of QDE-2 expression
by dsRNA is regulated at the transcriptional level, Northern
blot analysis was performed. As shown in Fig. 1D, the ad-
dition of QA led to a significant increase in qde-2 mRNA
levels in strains containing the dsRNA constructs, indicating
that dsRNA increases QDE-2 expression at the transcriptional
level. Although the qa-2 promoter is tightly regulated by the
presence of QA, it is not a very strong promoter (13). The
dramatic inductions of qde-2 mRNA and QDE-2 protein in
the presence of QA in the dsRNA construct-containing strains
suggest that the amount of dsRNA produced endogenously
is low.
dsRNA, not siRNA, induces QDE-2 expression, and DCLs
are required for the maintenance of QDE-2 levels posttran-
scriptionally. The low levels of QDE-2 in the dcl-1kodcl-2rip
double mutant (Fig. 1A) suggest that DCLs are required for
the maintenance of steady-state levels of QDE-2. Further sup-
porting this notion, we found that the induction of dsRNA in
the dcl-1kodcl-2ripdouble mutant failed to increase the level of
QDE-2 (Fig. 2A). In contrast, QDE-1 was not required for the
induction of QDE-2 by dsRNA. It has been previously shown
that QDE-1 is not required for gene silencing when dsRNA is
produced from an exogenous hairpin construct (10).
The data presented above suggest that the DCLs and siRNA
either regulate QDE-2 posttranscriptionally or are required for
FIG. 1. Induction of qde-2 expression by dsRNA expression.
(A) Western blot analysis showing that the levels of QDE-2 are low in
the RNAi mutants. The asterisk indicates a nonspecific cross-reacting
protein band recognized by our QDE-2 antibody. Equal protein load-
ing of the gel was ensured by the amido black-stained membrane
shown below the blot. Liquid cultures were grown for 2 days before
harvesting. (B) Western blot analysis showing the levels of QDE-2 in
the wild-type (WT) and al-1-quelled strains. (C and D) Western blot
(C) and Northern blot (D) analyses showing the induction of the
QDE-2 protein and qde-2 mRNA, respectively, in strains with dsRNA
constructs. The liquid cultures were grown for 2 days (with/without
QA) before harvesting.
VOL. 27, 2007 DOUBLE-STRANDED RNA INDUCED RNAi 3997
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the transcriptional activation of qde-2. To distinguish between
these two possibilities, Northern blot analyses were performed
to examine the expression of qde-2 mRNA after the induction
of dsRNA in the dcl-1kodcl-2ripdouble mutant (Fig. 2B). We
found that the transcriptional activation of qde-2 by dsRNA
was maintained in the dcl-1kodcl-2ripdouble mutant. In the
wild-type strain, qde-2 mRNA levels were induced after 1 h,
peaked 4 to 6 h after the addition of QA, and decreased
afterwards. In comparison, qde-2 was activated to a higher level
in the mutant, and high levels of qde-2 mRNA were maintained
12 h after the addition of QA. In addition, the basal level of
qde-2 was higher in the mutant than in the wild-type strain.
Despite the increase in qde-2 mRNA in the dcl-1kodcl-2rip
double mutant, QDE-2 levels failed to increase significantly. In
contrast, in the wild-type strain, the increase of the QDE-2
protein was seen 1 to 2 h after the QA treatment; QDE-2 levels
were maintained at high levels (?10-fold of the basal level)
after 8 h. Since DCL-1 and DCL-2 are responsible for all Dicer
activity in Neurospora (10), these data indicated that dsRNA,
but not siRNA, is responsible for the transcriptional activation
of qde-2 expression, and DCLs are not required for dsRNA
sensing in the qde-2 transcriptional activation pathway. How-
ever, DCLs are required for QDE-2 accumulation posttran-
scriptionally, suggesting that siRNA, the cleavage product of
dsRNA by DCL, plays a role in the accumulation of the
QDE-2 protein.
Induction of DCL-2 by dsRNA. The induction of qde-2 ex-
pression by dsRNA prompted us to examine whether the other
key component of the RNAi pathway, DCL-2, is also regulated
by dsRNA. Although the functions of DCL-1 and DCL-2 are
partially redundant, DCL-2 is responsible for more than 90%
of the Neurospora Dicer activity (10). As shown in Fig. 3A,
dcl-2 mRNA was strongly induced by the production of dsRNA
in two strains with different dsRNA constructs, indicating that
dsRNA also transcriptionally activates dcl-2 expression. Exam-
ination of the DCL-2 protein showed that the production of
dsRNA also led to a significant increase in DCL-2 levels (Fig.
3B). In contrast, the presence of QA had no effect on dcl-2 or
DCL-2 levels in the wild-type strain. Interestingly, the kinetics
of DCL-2 induction by dsRNA were significantly delayed com-
pared to those of QDE-2; DCL-2 levels did not peak until 24 h
after the addition of QA. These data suggest that the induction
of dcl-2 by dsRNA is a secondary response rather than an
immediate response as presumed for qde-2.
To understand whether QDE-2 participates in the signaling
pathway that mediates the induction of gene expression by
dsRNA, we examined the induction of DCL-2 by dsRNA in a
qde-2 null strain (qde-2rip). As shown in Fig. 3C, the induction
of DCL-2 by dsRNA was normal in the qde-2ripstrain. To-
gether with the data presented in the above-described figures,
FIG. 2. DCLs posttranscriptionally regulate the steady-state levels
of QDE-2, and dsRNA, but not siRNA, is responsible for the tran-
scriptional activation of qde-2. (A) Western blot analysis showing that
the level of QDE-2 could not be induced by dsRNA in the dcl double
mutant. The liquid cultures were grown for 2 days (with/without QA)
before harvesting. (B) Northern and Western blot analyses showing
the induction of qde-2 expression by dsRNA in the wild-type (wt) and
dcl double mutant strains. The number of hours indicates the time
after the addition of QA. The asterisk indicates a nonspecific cross-
reacting protein band recognized by our QDE-2 antibody.
FIG. 3. Induction of dcl-2 expression by dsRNA. (A) Northern blot
analysis showing the induction of dcl-2 mRNA in strains with dsRNA
constructs. (B) Western blot analysis showing the induction of DCL-2
protein by dsRNA. The number of hours indicates the time after the
addition of QA. The same membrane used in Fig. 2B was used here.
(C) Western blot analysis showing that the induction of DCL-2 by
dsRNA is normal in the qde-2 mutant strain. The asterisk indicates a
nonspecific cross-reacting protein band recognized by our DCL-2 an-
tibody. WT, wild type.
Page 5
these data demonstrate that the components of the RNAi
pathway are induced by dsRNA but are not required for
dsRNA sensing and transcriptional activation in Neurospora.
Induction of QDE-2 by dsRNA is required for optimal gene
silencing. Since QDE-2 and DCL-2 are two central players in
the RNAi pathway, the induction of QDE-2 and DCL-2 by
dsRNA suggests that their regulation is important for the ef-
ficiency of the pathway. To test this hypothesis, we examined
whether the high QDE-2 level induced by dsRNA is required
for the efficiency of gene silencing. A construct in which the
qde-2 is under the control of the qa-2 promoter (qaQDE-2)
and a construct containing the wild-type qde-2 gene with its
own promoter were created and transformed into the qde-2rip
dsal-1 strain. As shown in Fig. 4A, the wild-type qde-2 con-
struct complemented the function of the endogenous qde-2,
and the transformant exhibited dsRNA-induced QDE-2 ex-
pression (Fig. 4A) and silencing of the al-1 gene, as indicated
by the white aerial hyphae and conidia in the presence of QA
(Fig. 4B). On the other hand, the expression levels of QDE-2
in qaQDE-2 transformants in the presence of QA were com-
parable to the uninduced level of QDE-2 in the qde-2ripdsal-1
qde-2 strain. In addition, the qde-2ripdsal-1 qaQDE-2 strains
showed only very weak silencing of al-1 in the presence of QA,
indicating a severe compromise of RNAi efficiency. These re-
sults suggest that the high levels of QDE-2 induced by dsRNA
are required for efficient RNAi and that the induction of
QDE-2 expression by dsRNA is mediated by the qde-2 pro-
To further confirm this result, we created constructs that
contain both the qde-2 gene with segments of its upstream
sequence of different lengths (Fig. 4C) and the dsal-1 cassette.
These constructs were transformed into the qde-2ripstrain at
the his-3 locus. As shown in Fig. 4D, Pqde-2A, which contains
1.9 kb of qde-2 upstream sequence, fully complemented the
function of the endogenous qde-2, as indicated by the induction
of QDE-2 and silencing of al-1 in the presence of QA in the
transformants. In contrast, in the Pqde-2B transformants,
dsRNA-induced QDE-2 expression was abolished; the QDE-2
levels remained at the basal level in the presence of QA. These
data indicate that a cis element within the 0.4-kb qde-2 pro-
moter is required for dsRNA-induced expression. Also, as ex-
pected, the aerial hyphae and conidia of the Pqde-2B strains
remained orange in the presence of QA, indicating a severe
compromise of the RNAi pathway. Together, these data indi-
cate that QDE-2 is a limiting factor in the RNAi pathway and
that the induction of QDE-2 by dsRNA at the transcriptional
level is critical for RNAi function.
Genome-wide search revealed that additional RNAi compo-
nents and genes homologous to host defense responses are
DRAGs. The observation that qde-2 and dcl-2 were induced by
dsRNA indicates the existence of a transcription-based dsRNA
response program in Neurospora. To understand the function
of this response, we carried out a genome-wide study to iden-
tify other DRAGs in Neurospora by microarray and qRT-PCR
analyses. To identify genes that are immediately activated by
dsRNA, cultures of the dsal-1 strain were treated with QA for
6 h. The wild-type cultures treated with QA were used as the
controls. As shown in Fig. 5A, as expected, qde-2 mRNA was
dramatically induced in the dsal-1 cultures. In contrast, the
qRT-PCR analysis showed that the levels of alpha-actin and
qa-2 (a QA-inducible gene) mRNAs were comparable in both
strains (Fig. 5B), indicating that the QA responses were similar
in both strains. Thus, activation of genes in the dsal-1 strain
should be due to the production of dsRNA. In addition to the
microarray experiments, genes known to be involved in the RNAi
machinery and some genes with low signal levels in the microar-
ray experiments were analyzed by qRT-PCR.
The microarray experiments showed that the expression of
FIG. 4. Induction of QDE-2 by dsRNA is required for efficient
RNAi. (A) Western blot analysis showing the expression of QDE-2 in
the indicated strains. (B) Photograph of the corresponding strains
growing in slants. (C) Graphic depiction of the indicated qde-2 pro-
moter constructs. (D, top) Western blot analysis showing the expres-
sion of QDE-2 in the indicated strains. (Bottom) Photograph of the
corresponding strains growing in slants. The asterisk indicates a non-
specific cross-reacting protein band recognized by our QDE-2 anti-
body. wt, wild type.
VOL. 27, 2007DOUBLE-STRANDED RNA INDUCED RNAi 3999


text:
Redbridge Jewish Care A remain the only team in the JC MSFL Premier Division with a 100 per cent record after they beat London Maccabi Lions A 2-1. Ben Sollosi and Lewis Walker were their scorers. Mike Pearson pulled one back for the Lions.
Two goals from Jacob Gold helped North London Raiders A end Faithfold A's perfect record in a 3-2 victory. Rob Sanford made it 3-0 before Paul Babai and substitute David Lederman set up a grandstand finish.
A new seven-a-side Jewish football league is kicking off in Hendon on Sunday afternoons from October.
The friendly league, which is being run by Hendon United Sports Club, is open to all and designed to appeal to both children and adults who do not want to play in formal league teams.
Anyone from six upwards can make up teams, existing clubs can enter teams, or individuals can enter, in which case the league will help them form teams.
Hyde Park Rovers boss Phillip Braham has warned his hot-headed students to leave their raging antics at home following their fiery clash against Manchester Maccabi 3rds.
Rovers were reduced to nine men after having Bobby Abrahams and Jason Wise sent off during their hard-earned 2-1 victory over the previously unbeaten league leaders.
Both were dismissed for two bookable offences as tempers flared in a pulsating contest which saw Martin Simons' men suffer their first defeat in six games.
Yossi Benayoun is set to undergo the controversial horse placenta treatment from Serbian physiotherapist Marijana Kovicevic in a bid to be fit for Israel's upcoming vital Euro 2012 qualifiers.
The Chelsea midfielder limped off in the second half of last week's Carling Cup defeat against Newcastle last week after tearing a muscle in his right calf. The initial diagnosis was that he would be out for at least three weeks, missing Israel's home game against Croatia on October 9 and visit to Greece on October 12.
London Lions took a valuable point against high flying Hoddesdon Town in the South Midlands League Division One.
Andy Glynne's second-half goal ensured Lions took a point after a poor first-half performance.
Both sides were devoid of regular first team players and the game and result reflected that. Among the absentees for the Lions were top-scorer James Gershfield, Adam Stolerman, Craig Ellis, Danny Berg and Ben Ellis. Blustery conditions in Broxbourne made it difficult for both teams to find any rhythm and resulted in a scrappy affair.
Uefa have announced that Howard Webb will officiate Hapoel Tel Aviv's Champions League Group Stage match with Lyon at Bloomfield Stadium on Wednesday night.
Webb, 39, is considered the world's top referee after taking charge of the World Cup final between Spain and Holland in South Africa in July and last May's Champions League final between Inter Milan and Bayern Munich.
Webb took charge of the goalless Premiership draw between Fulham and Everton at Craven Cottage on Saturday.
Hapoel lost their opening Group B match 2-0 against Benfica. They currently lie 8th in Ligat Ha'al.
Manchester Maccabi 1st progressed into the next round of the County Manchester County Cup after beating Middleton Colts 3-2. Adam Lavin, Michael Sacks and Rafi Leeker were the marksmen.
A goal from Andy Glynne helped London Lions come from behind to draw against Hoddesdon Town in the South Midlands League Division One.
Manchester Maccabi 1st progressed into the next round of the County Manchester County Cup after beating Middleton Colts 3-2.
In a re-arranged tie, it was colts who settled first, but a couple of long range shots did not really trouble Dan Garman in the Maccabi goal
FC Team moved into the third round of the Middlesex County Senior Open Cup after despatching Brentham FC 2-0.
Mitch Young's men arrived with a depleted squad with no substitutes and without a recognised keeper. Down to the bare bones, Young was forced to fill in between the sticks.
Yuval Spungin faces a backlash in Israel after playing for Omonia Nicosia in Cyprus in their 4-0 away win over Larnaca on Yom Kippur.
Spungin, a former Maccabi Tel Aviv defender, claimed he was compelled to play by his club.
Daryl Phillips, assistant-manager of Faithfold A, has called for calm after the Superhoops went three points clear at the top of the Premier Division after beating defending champions North West Neasden A.
Victories in their first three matches against title rivals Hendon United Sports Club A, London Maccabi Lions A and Neasden have put Faithfold in a strong position as they look to win the title for the first time since 2005.
Phillips said: "This was another squad performance where everyone did a job."

subset: commoncrawl


text:
Former WWE Divas Champion and TNA Knockouts Champion Mickie James talks about appearing at WrestleCon 2015 on March 27 and 28 in San Jose, being around her friends including Ric Flair, being a mom now and a wrestler, how she looks at her career now going forward, how close she came to retiring from the ring, her recent return to ...Read More »
Tag Archives: tna wrestling
The Flagship: Roman Reigns to Main Event WrestleMania 31, WWE Raw, Fast Lane and More
It's The Flagship podcast as Brian and Michael talk about WWE Fast Lane from Sunday night as well as Raw from Monday, Roman Reigns to main event WrestleMania 31 against Brock Lesnar and the response from the audience towards him lately, John Cena vs. Rusev heating up and ideas for the buildup to their match and lots more. Plus, they ...Read More »
Scott Fishman Talks NXT Brand, Fast Lane, Samoa Joe Leaving TNA and More
Scott Fishman – contributor to the Miami Herald fighting section – joins the podcast to talk about the NXT brand and where it's at right now, what the WWE could learn from how that brand is booked, possible callups, the joy of house shows, Fast Lane on Sunday and how it could be sneaky good, thoughts on where they are going ...Read More »
Accentuate the Positives in Wrestling – Week of February 14, 2015
It's easy to get bogged down in the negativity that exists online when it comes to the world of professional wrestling (or "Sports Entertainment" if you prefer). Are there things worthy of that negativity in the wrestling world? Absolutely! But too often, there are positive aspects to the current product that get overlooked. That's what I'm aiming to correct with ...Read More »
The Flagship: Daniel Bryan and Roman Reigns are Mad, Seth Rollins May be Sad and NXT Talk
It's this week's edition of The Flagship as Brian and Michael cover the week in wrestling including the Seth Rollins photo leak, Raw from Monday night and what is going on with Daniel Bryan and Roman Reigns, the Triple H / Sting angle, the Triple H conference call with the media including his plans for NXT in 2015, the NXT ...Read More »
The Flagship: Road to WrestleMania in Question with Roman Reigns and Daniel Bryan, Triple H Talks with Stone Cold
Brian and Michael talk about WWE Raw from Monday night including the possible changes on the Road to WrestleMania involving Roman Reigns and Daniel Bryan as they two are set to square off at Fast Lane, if WWE make a switch for the main event of WrestleMania 31 and who will face Brock Lesnar, protecting Reigns and how tricky that ...Read More »
Scott Fishman Talks Randy Savage Induction, Daniel Bryan Return and Magic City Con
Scott Fishman — contributor to the Miami Herald fighting section — is back on the podcast as he talks about the Magic City Con this weekend down in Miami and the various wrestlers appearing he has spoken to, "Macho Man" Randy Savage being finally being inducted into the WWE Hall of Fame, where he once met Savage, his recent interview ...Read More »
The Flagship: Randy Savage going into WWE Hall of Fame, Raw Fallout, TNA Debut
It's The Flagship podcast with Brian and Michael and there's a lot to talk about! The guys discuss "Macho Man" Randy Savage finally going into the WWE Hall of Fame and the influence he had on the business along with who else might be joining him as part of the Class of 2015. Plus, they talk about Raw from Monday ...Read More »
Mauro Ranallo Talks New Japan Pro Wrestling on AXS TV, His Style Calling the Action and the Wrestling Landscape
Combat sports announcer Mauro Ranallo joins the podcast to talk about New Japan Pro Wrestling coming to AXS TV with the 13-epidsode series premiering on Friday, Jan. 16 at 9pm ET, his return to calling professional wrestling, what has always drawn him to NJPW, teaming with Josh Barnett to call the shows and what the sessions were like, if he ...Read More »
The Flagship: The Authority is Back, TNA Wrestling is Back on TV, Wrestle Kingdom 9 Recap
It's The Flagship podcast with Brian and Michael as they talk about TNA Wrestling moving to its new home on Destination America, expectations for them there, Brian's recent interview with Dixie Carter and comments she made there. Plus, they talk about WWE Raw from Monday night which was another lacklusted edition even with The Authority back in power along with ...Read More »
Best of Wrestling 2014: Daniel Bryan Wins at WrestleMania, Seth Rollins Emerges, The Streak is Over
Brian and Michael look back at the year in wrestling in the best of wrestling 2014 podcast. The guys run down who they thoughts was the best wrestler, heel and tag-team over the past year along with their best match and show. Plus, they talk about this biggest misfire along with the biggest news story as well as what they thought ...Read More »
The Flagship: WWE Raw, Daniel Bryan is Back, What Lies Ahead
It's The Flagship podcast with Brian and Michael as they talk about WWE Raw from Monday night including the return of Daniel Bryan, what his coming back could mean for plans going forward and into WrestleMania 31, the return of The Authority, several guys getting chances to do promos during the show, Seth Rollins changes of winning the WWE title ...Read More »


text:
It's time to find out whether Toshiba's debut Ultra HD TV can keep up with the 4K Joneses
World's first HDMI 2.0 Ultra HD TV throws open the doors to a high frame-rate future
Immense black levels, immersive 3D and punchy colours at an attainable price point
Superb 55in LED TV makes Ultra HD tech more manageably sized and affordable than ever
While this 55in flagship Panasonic LED is a real looker, it's not without some flaws
LG's gorgeous-looking TV comes close to greatness, but suffers in regards to contrast
Sony's Triluminos technology ensures its 55in W9 model packs a real picture punch
Can this whopping 85in Ultra HD TV really justify its equally mammoth £35,000 price tag?
Philips' topline Full HD set offers extensive processing tweaks - and you'll need to master them
There's more to Samsung's 8000 Series LED than its glam design and Smart talents
The resolution revolution begins as Sony brings 4K to the masses with this 65in Ultra HD TV
Tech brand's 'step-down' F7000 series LED TV isn't that much of a step down after all
Get connected for less with this sub-£1,000 42in Smart 3DTV
Plasma TV tech may well have reached its zenith with the ZT65. If only it wasn't a limited edition...
Brand's largest consumer plasma is a drop-dead gorgeous edifice with a features list to die for
Forget gesture control and speech recognition, this flatscreen flagship is all about image quality
B&O's new 40in set startles with its AV performance and its more-than-£5,000 price tag...
It's as stylish as LED TVs get - but is there more to this flagship screen than its sultry design?
Active 3D and multimedia tricks for only £450 - but this 40in LED TV isn't without its flaws
This leading-edge 47in LED TV offers more control and customisation than any set before it
Plasma TV tech may well have reached its zenith with the ZT65. If only it wasn't a limited edition...
Razor-sharp pictures and a revamped Smart system ensure this mid-range 50-incher is ready to thrill
This TV wants to be a soundbar-slayer. And in that regard it succeeds
This big, curved 4K TV impress in every way
Flagship 65in 4K/Ultra HD TV remembers that audio is just as important as picture quality
Love Home Cinema? Sign up to our emails for the latest news and special offers!
Ultra HD for all: Five sub-£1,000 Ultra HD TVs fight it out in this month's grouptest
Hot Technology: Ultra HD Blu-ray, OLED TVs and more from the IFA tech expo
Top flight TV: Hands-on with Panasonic's best LED screen yet!
Sonic sensations: The Dolby Atmos discs you must own
Win! Win! Win! Three Evolve Audio SB-2501 soundbars up for grabs!
Plus: All of the latest home cinema tech,
Blu-ray/DVD reviews, and a whole lot more!
Want to see your home cinema system featured in the pages of HCC? Click here for more info.
Home Cinema Choice is proud to be a member of EISA.
Visit www.eisa.eu for more info.


text:
Company Overview of Window to the World Communications, Inc.
Executive Profile
Renée Crown
Vice Chairman and Member of 98.7 WFMT & The WFMT Radio Network Committee, Window to the World Communications, Inc.
|Total Calculated Compensation
|This person is connected to 3 Board Members in 3 different organizations across 3 different industries.
See Board Relationships
Mrs. Renée Crown serves as a Vice Chairman of Corporate Board for Boys and Girls Clubs Of Chicago. Mrs. Crown serves as a Vice Chairman at Window to the World Communications, Inc.
Corporate Headquarters
5400 North Saint Louis AvenuePhone: 773-583-5000
Chicago, Illinois 60625-4698
United States
Fax: 773-509-5303
Board Members Memberships
Vice Chairman of Corporate Board
Vice Chairman and Member of 98.7 WFMT & The WFMT Radio Network Committee
EducationThere is no Education data available.
Other Affiliations
Annual CompensationThere is no Annual Compensation data available.
Stocks OptionsThere is no Stock Options data available.
Total CompensationThere is no Total Compensation data available.
|There is no Competitor Compensation data available.
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text:
Jennifer Lansford is looking at the ways parents express love for their children. The data -- gathered by studying parent-child relationships in the U.S., China, Colombia, Italy, Jordan, Kenya, Philippines, Thailand and Sweden -- reveal insightful differences in the ways love is expressed.
Read Whole Story
In Tokyo, a single mother who works as an editor for HuffPost Japan rises at 5 a.m. each workday to pack a lunch for her teenage son before leaving fo...
Every society values its own skill sets for its own reasons. The problem in America is that, while we excel at raising spectacularly verbal kids, we have lost sight of other values that we need to foster in our children.
Though parents around the world have the same goals, American parents like me (despite our very best intentions) have gotten it all backwards.
After working in England, Germany and the United States and hearing from friends who live all over the world, I've noticed a few places where modern American parents could use a bit of extra coaching.
It was one of the eye-opening experiences inspired by my years living abroad raising my children and researching parenting beliefs and practices in different cultures. There were so many wow moments, when my middle-class mom instincts clashed with what I was living, hearing, seeing and reading.
Get top stories and blog posts emailed to me each day. Newsletters may offer personalized content or advertisements. Learn more.


text:
Who will be elected to City Hall? Here's a glance at the candidates running in each City Council district in the May 11, 2013 election.
San Antonio Express-News and Courtesy Photos
DISTRICT 7 CANDIDATES, CLOCKWISE FROM TOP LEFT: Incumbent Cris Medina, Michael A. Gonzales, Gloria Rodriguez and Carlos Zaldivar. Click here to read more about the District 7 race


text:
Homes & Houses for Sale ListingsStaten Island, NY
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$85,000,000 - 10 Bed, 13 Bath
River 2 River Realty
New York,NY10036
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$80,000,000/8 Bed, 10 Bath
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New York,NY10004
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Zeckendorf Marketing LLC
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$70,000,000/4 Bed, 7 Bath
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New York,NY10012
$66,000,000/5 Bed, 7 Bath
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Extell Marketing Group
New York,NY10065
$65,000,000/6 Bed, 7 Bath
The ATELIER SALES OFFICE - Call us at 212-563-6635 for an immediate appointment. A 2 million dollar construction credit as well as a 1M Yacht with docking fees for 5 years will be included in the sa...
Built in 1901 and designed by the firm of Welch, Smith & Provot, this exceptionally grand and remarkably beautiful house was constructed in the Beaux-Arts style and is rightfully regarded as an archit...
Perched high atop The Residences at the Ritz Carlton, the opportunity exists to create a truly unrivaled masterpiece duplex Penthouse with sweeping 360 degree views of New York's most important landma...
Perfectly positioned high above Madison Square Park and crowning the One Madison Condominium, this grand-scale Penthouse with nearly 7,600 square feet of interior and terraced luxury has been designed...
The Duplex Penthouse is an incredible home in the sky, surpassing in size, breadth, light and views anything that has previously existed on the Upper East Side. A distinct separation between public an...
The crown jewel of Chelsea's most important address - Walker Tower's Penthouse One is beyond description, it's to be experienced. Please contact Kamali Chandler for additional details or to schedule ...
This spectacular 16-room duplex apartment is situated high atop the prestigious Pierre Hotel on Fifth Avenue. Featuring a meticulous triple mint renovation and boasting extraordinary 360 degree views ...
Discover a breathtaking world of privacy and prestige, the Duplex Penthouse at 50 United Nations Plaza, the ultimate global address. Designed by Pritzker Prize winning architects Foster + Partners an...
3.5 Co-Broke Commission to Buyer's Broker. First Offering. The Crown Jewel of SoHo. Introducing Penthouse I, a newly completed, sprawling duplex condominium atop the famed Puck Building overl...
This five story newly constructed luxury Town House offers nearly 10,000 SF of interior space and over 2,500 SF of outdoor space. This exclusive home features a beautiful limestone facade and a separa...
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text:
|Project Name
|Price (USD)
|Project Name
|Price (USD)
|Flash Action script Mp3 Player
|Need a custom flash mp3 player that will read from an xml file. Heres an example of how I would like...like it to look. http://www.podsnack.com/mp3-player-examples/players-with-playlists#2 See attached
|PHP, Adobe Flash, Graphic Design, ActionScript
|Apr 1, 2012
|Apr 1, 2012Ended
|mp3 Player - Some action scripting
|I need some action script programming to a given .fla file. All layouts are allready done. Just some...some simple mp3 functionality should be added. ## Deliverables 1) Complete and fully-functional working
|PHP, Javascript, Adobe Flash, Engineering, Software Architecture, Software Testing, Website Testing, MySQL, Website Management, Web Hosting
|Feb 22, 2007
|Feb 22, 2007Ended
|Simple action scripting - mp3 player
|I need some action script to a existing .fla file. All layouting is allready done. This should be an...an simple mp3 player. ## Deliverables 1) Complete and fully-functional working program(s) in executable
|PHP, Javascript, Adobe Flash, Engineering, Software Architecture, Software Testing, Website Testing, MySQL, Website Management, Web Hosting
|Feb 22, 2007
|Feb 22, 2007Ended
|Need urgent fix two action script issues on mp3 player
|I want to fix two action script issues for my mp3 player urgently.Fla file provided and more detail discuss... This is max 2 hours work for experienced action script provider. Budget $30. Thanks Tom
|XML, Adobe Flash, ActionScript
|Apr 16, 2010
|Apr 16, 2010Ended
|165373 Simple Flash Mp3 Action Script
|Simple Flash MP3 Player Button Action Script: --------------------------------------------- 1. I...and stop play of a single mp3 file. Variables should be set for 1. Mp3 file name 2. Auto play
|XML, Adobe Flash, Website Design, Anything Goes, HTML
|Sep 13, 2007
|Sep 13, 2007Ended
|Flash player for wav and mp3
|Hi, We need a flash player that can play the following audio format: mp3, wav It must support all...with no distortion. It should support multiple player on the same web page, which when clicking play... The player should have: play, stop, volume and progress bar, with option for mini player with play/stop
|Adobe Flash
|Jun 26, 2013
|Jun 26, 2013Ended
|Simple Flash MP3-player
|We need a simple Flash MP3-player for our site. The only elements in the player is play-button and a stop-button...have to do is the Action Script coding. **Specifications:** * Flash MP3-player, with play and stop-buttons...stop-buttons * The player should be initialized with preview_player.swf?id=XXXX, and load the file
|Jul 6, 2010
|Jul 6, 2010Ended
|Timer on MP3 Flash Player
|...made flash 8 MP3 player to start counting up... showing (0.00 min.sec) as the music player plays. Needs...Needs to follow aready written Action Script found in the sound.fla which I will provide once we have an...click on the right menu "SOUNDTRACK" the music player functions as needed but notice the 00.00 needs
|PHP, Adobe Flash, Engineering, Software Architecture, Software Testing, Website Testing, MySQL, Website Management, Web Hosting
|Aug 9, 2006
|Aug 9, 2006Ended
|mp3 player (a)
|i'd like to have an mp3 player like here for my site: http://new.music.yahoo.com/charts/ click..."play" onto a specific song so you can see the player in action on the bottom left. I also need a scrubber
|Jul 11, 2008
|Jul 11, 2008Ended
|141174 Flash Player/Action Needed
|...Listening Experience for a potential Flash/Action Script contract. Instatone is a new community DIY radio...play songs 'on demand' via our opensource flash player it plays on top of the radio program that streams...streams via shoutcast. User has to go into default player (ie iTunes) and actually stop the stream as it
|Adobe Flash, Website Design, Anything Goes
|May 17, 2007
|May 17, 2007Ended
|football player action script
|Hi, You worked on an ActionScript project for me. Can you get back in touch as I am willing to pay for you to finish the work Thanks
|PHP, Javascript, Adobe Flash, Facebook Marketing, ActionScript,
|Feb 20, 2015
|Feb 20, 2015Ended
|Flash MP3 Radio Player (Advanced)
|Flash MP3 Player Project to include the Following - Graphic Animation as music plays - Embedded...respond if you are experienced with Flash and Action Script.
|Adobe Flash, ActionScript
|May 18, 2011
|May 18, 2011Ended
|35494 Flash Mp3 Player Modification
|...already built mp3 player. A simple job of combining the two files and adding some action script. I'd just...equalizer added to the stage and I'll fit it in the player later on. I'd like the start, stop and pause buttons...the equalizer to start as well. Here is the mp3 player as an swf http://ras-binghi.com/hear-reggae-music
|Adobe Flash, Anything Goes
|Feb 18, 2006
|Feb 18, 2006Ended
|Flow Player Action Script.
|We have to embed controls to Flow Player. As described on the attached document. ## Deliverables 1)...Worker Legal Agreement). ## Platform Flow Player, Linux
|Engineering, Software Architecture, Software Testing
|Jan 6, 2011
|Jan 6, 2011Ended
|Flash player for wav and mp3 - repost
|Hi, We need a flash player that can play the following audio format: mp3, wav It must support all...with no distortion. It should support multiple player on the same web page, which when clicking play... The player should have: play, stop, volume and progress bar, with option for mini player with play/stop
|Adobe Flash
|Jun 26, 2013
|Jun 26, 2013Ended
|mp3 player
|I need a simple falsh mp3 player similiar to npr.org or in.com where the player is in popup page. Provide...Provide me the script as a plugin for my wordpress site or in general php / javascript. Keep in mind...mind do not use jwplayer and player should not loose playlist information when main page is refreshed
|PHP, Javascript, Adobe Flash, Website Design, MySQL
|Aug 31, 2010
|Aug 31, 2010Ended
|Flash action script help
|...investigation that could work. 2) I want to load a mp3 player with a playlist (can it just be one of the pre-built
|PHP, Project Management, Engineering, Software Architecture, Software Testing, Website Testing, MySQL, ActionScript, Website Management, Web Hosting
|Oct 13, 2010
|Oct 13, 2010Ended
|249122 Flash MP3 Player Skin
|...new skin for a flash mp3 player. I can provide the .fla of the original player and also a .psd of what... I've attached 2 screen shots. One of what the player looks like now and one of what the new skin will...will look like. The functionality of the new player skin will be cut down a bit. For example not having
|Adobe Flash, Anything Goes
|Sep 5, 2008
|Sep 5, 2008Ended
|4704 Flash Mp3 Player Script Mod
|I have a flash mp3 player PHP script with an ecommerce backend that hooks up with Paypal to sell and...upload the mp3's and the paths are automatically written to the xml playlist (which the player reads) and...instant delivery. I'd like to add an option to this script. In the admin area I'd like the user to have an
|PHP, XML, Adobe Flash, MySQL, Anything Goes
|Aug 15, 2006
|Aug 15, 2006Ended
|Mp3 player
|I have a flash Mp3 Layout thats designed but I need the functionality of AS3 to pull the audio from an...XML file. The player has a next, previous, play, and pause button that loops 8 mp3 files. Below these...each Mp3 file individually. This script should be setup incase there would be more or less mp3 for
|Adobe Flash
|Aug 13, 2010
|Aug 13, 2010Ended


text:
30 results was found matching keyword
0 vote(s)Comment
VOV News English 2 week(s) ago
Australia's Fred Hollows Foundation has provided medical care for thousands of people with eye defects in the northern province of Thai Binh through its three-year project.
Keywords: CommunesprojectThai BinhBusiness OpportunitiesPoor Patients
VOV News English 3 week(s) ago
As many as 500 impoverished Cambodian patients will receive free health check-ups and treatment by Vietnamese and Cambodian doctors from September 10-12, raising the total number of beneficiaries this September to 1,000.
Keywords: Poor PatientsNguyen HoangCho RayBeneficiariescare forIn CambodiaDiscounts
VOV News English 2 month(s) ago
1 related news
A veteran in Ho Chi Minh City has devoted his life to providing free treatment for poor people coming from across Vietnam during the past 20 years.
Keywords: Golden HeartVan TamreluctantlyHoc MonLuckierPoor Patients
VietnamNet English 4 month(s) ago
VietNamNet Bridge – Vietnamese doctors have to decide whether they should sign personal guarantees for poor patients so they can be treated without having to wait for a deposit to be made.
Keywords: Poor PatientsguaranteeDoctorssigndoctorcap
Báo Tuổi Trẻ English 5 month(s) ago
2 related news
The Orbis Flying Eye Hospital carrying experts from the UK, France, the U.S., and Canada landed in the central city of Hue on May 5 to start its two-week program to offer free surgery and health checks to poor patients.
Keywords: ChecksPoor PatientsHue Central HospitalFacebookTwitteroffer
VietnamPlus English 7 month(s) ago 1 duplicate news
The Ho Chi Minh City Dermatology Hospital together with its French partner inaugurated a new leprosy treatment facility on March 4.
Keywords: Health DepartmentCertificatesfrenchjustifyconsolidatePoor PatientsA French
Báo Dân Trí English 10 month(s) ago
Doctors and patients at Cho Ray Hospital in HCM City have called for tighter controls over charity groups that provide food to poor patients after an incident in which rotten meals were served.
Keywords: Food ToImagesCho Ray HospitalThanh Hungdailydonate
VietnamPlus English 11 month(s) ago
1 related news
As many as 400 poor patients in the Mekong Delta province of Dong Thap's Cao Lanh city have been provided with free medical check-up by doctors from the provincial General Hospital as part of the 2014 community healthcare campaign.
Keywords: Dong ThapcheckCao Lanh CityjustifyPoor PatientsGeneral Hospital
Báo Tuổi Trẻ English 12 month(s) ago
1 related news
Around 80 poor patients received free operations and joint replacements at a hospital in Hanoi from October 9 to 12 thanks to the support of local and American doctors from the US-based Operation Walk.
Keywords: Operation WalkThe U SDoctorsgroinBut VietnameseCentral Hospital
Báo Thanh Niên English 12 month(s) ago
1 related news
Around 80 poor Vietnamese patients received free knee and hip replacements from the US's Operation Walk between September 9 and 12 in Hanoi.
Keywords: Operation WalkvolunteerdatePoor PatientsConcernsChoosing
VietNam News English 13 month(s) ago
As much as 70 per cent of breast cancer patients are diagnosed when they are already in the final stages of the disease, said Deputy Minister of Health, Nguyen Thi Xuyen at a recent breast cancer prevention event in Ha Noi.
Keywords: StageslateHealth Nguyen Thi XuyenPoor Patientscomehelp
VOV News English 14 month(s) ago
The Ho Chi Minh City branch of the Supportive Fund for Cancer Patients – the Bright Future was launched on September 9 at the Oncology Hospital.
Keywords: DisadvantagedbranchPoor PatientsThe CostsrefuseDanangThe Ho Chi Minh City
VOV News English 14 month(s) ago
1 related news
(VOV) - Cataract surgery brings about high economic efficiency to Vietnam, according to a latest research VISONARY conducted by Fred Hollows Foundation (FHF) and the George Institute for Global Health.
Keywords: visuallyPoor PatientsVietnam AccordingrestorePushingbetter
(VOV) - Cataract surgery brings about high economic efficiency for Vietnam, according to the latest research VISONARY conducted by Fred Hollows Foundation (FHF) and the George Institute for Global Health.
Keywords: Do Nhu HonvisuallyPoor PatientsFor VietnamVietnam Accordingrestore
VOV News English 15 month(s) ago
1 related news
(VOV) -The Prudence Vietnam Foundation on August 5 donated VND600 million to the National Fund for Vietnamese Children (NFVC) to provide free eye surgeries for 200 needy and disabled children.
Keywords: visuallyPrudential VietnamPoor PatientsVietnamese ChildrenViettelprovide
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Helen Clark became the Administrator of the United Nations Development Programme in April 2009, and is the first woman to lead the organization. "I can think of no better way to spark conversations around the world on development issues than through the dynamic use of social media," said Helen Clark in reaction to her recent listing in the UK newspaper, The Guardian, as the tenth most interesting person tweeting about global development.
International Women's Day 2014 Twitter Chat
Join our administrator Helen Clark live on Twitter and ask her all your questions about women's empowerment, gender equality and how equality for women means progress for all!
Use the hashtag #UNDPchat in your tweets to join.
Past Chats with Helen Clark
Twitter chat with Helen Clark on HIV and Development
Thanks to all of you for joining our twitter chat about HIV and Global Development with United Nations Development Chief Helen Clark! In case you missed it - here's a selection of tweets from the chat. Continue the conversation during @aids2012 and use #AIDS2012 in your tweets. more
Twitter chat with Helen Clark on Sahel Crisis
Food security will become a major issue in the Sahel during the coming months. Helen Clark and Valerie Amos visited the Tillabery region of Niger on a two day mission, to highlight the looming crisis and emphasize the need for long term solutions in the region. more
@HelenClarkUNDP - #1 Goal is 2 Eradicate Xtreme Poverty
New York - My number one goal as Administrator of UNDP is to work for the elimination of extreme poverty, said United Nations top development chief more
UNDP chief tweets for women
New York - UNDP chief Helen Clark celebrated International Women's Day with a live Twitter chat from UNDP headquarters today in New York City. "I more
"Desire for Dignity" Driver of Change in Arab World - Helen Clark
New York — In her second Twitter chat, UNDP Administrator Helen Clark answered questions from around the globe on development in the Arab world. The questions ranged more
Our Actions Against Poverty
Related Links
- Momentum on the MDGs and Gender Equality in Haiti: A Conversation with UNDP and the Better World Campaign
- 2013 Human Development Report - Twitterchat with Helen Clark
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- Helen Clark's Interactive Lectures at Cambridge University
- Facebook Chat on International Women's Day with Rebeca Grynspan
- Social Media: an Outside the UN Perspective
- Storify | Social Media - An outside the UN perspective
- Twitter Chat on the Launch of the 2011 Human Development Report
- Video: Backstage at the Twitter chat with Helen Clark
- Twitter Chat on Arab Spring
- Twitter chat on International Day for the Eradication of Poverty
- Twitter Chat on International Women's Day
- Become our Digital Agent of Change
About Helen Clark
Our Perspective
27 Apr 2015:Le lexique électoral du PNUD brise les barrières linguistiques
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- Meet Archippus T. Sesay from Sierra Leone. He just won the 2015 Youth Innovation Award. An amputee himself, after seeing colleagues struggle with poor quality prosthetics, Achippus developed prosthetics using local materials and discarded waste that are 10 times cheaper than imported prosthetics and are just as durable: 13 hours ago
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Amore Opera Presents LA BOHEME, 12/14
This Winter, Amore Opera revives its classic production of Puccini's timeless masterpiece La Bohème. This production launched the company's inaugural season, and its revival promises to be equally riveting. La Boheme is an opera in four acts, with libretto by Luigi Illica. Based on Scènes de la Vie de Bohème by Henri Murger, it is the story of a group of young bohemians living in the Latin Quarter of Paris in the 1840's, and the tragic tale of two lovers doomed by circumstance and illness. La Bohème is fully staged with full orchestra and presented in its original Italian (with subtitles).
Amore Opera's production will be directed once again by Nathan Hull, and Gregory Ortega, who led the orchestra for last season's production of La Traviata, will conduct. Maestro Ortega has conducted with most of the major small opera companies in New York City, Boston and Florida, including New Rochelle Opera, Treasure Coast Opera and New York Concert Artists, in addition to having founded The Hunter College Opera Theater. Director Nathan Hull staged this season's Don Pasquale and the critically acclaimed American Debut of Olivo e Pasquale. He also directed last season's successful American premiere of I due Figaro.
Amore Opera will round out its winter offering with an "Opera-in-Brief" presentation of La Bohème. These ever popular, 90-minute performances of abridged works are geared towards young audiences and families, and are a great way to introduce children to the splendor of operatic performances. (Please note that due to the Hurricane, the production of "Scrooge & Gilbert & Sullivan" has been delayed until Dec. 2013).
Additionally, Amore will host its annual New Year's Eve gala. In addition to a ticket to the evening's performance of La Bohème, the gala includes hors d'oeuvres, dinner between acts, a champagne toast at midnight, and a special post-midnight concert.
All performances are at the Connelly Theater (220 East 4th Street), located on the South Side of East 4th Street between Avenues A & B. The closest subway is the F train at 2nd Avenue-Houston Street, 1st Avenue exit.
General admission is $40.00 and $30.00 for students and seniors. Tickets for the New Year's Eve Gala are $125. All tickets for the Opera-in-Briefs are $15.00.
Tickets can be purchased directly on the Amore Opera website at www.amoreopera.org or by calling 1-888- 811-4111.
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Melva Ellen
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Apr. 11, 1903
Schuylkill County
Pennsylvania, USA
Oct. 19, 1977
Lehigh County
Pennsylvania, USA
Born in the coal-region of McAdoo, Schuylkill County, Pennsylvania, Melva Ellen Helfrich was a daughter of William H Helfrich and
Cora Ellen (Schollenberger) Helfrich
She was sister to Grace, Harold, and Ellsworth Helfrich. She also had a brother who passed young, Claud Morton Helfrich.
Sometime in her teens, her (apparently "upwardly mobile") mother declared "I will not have my daughters marrying coal-crackers" and persuaded their father to move the family to Allentown, Lehigh County, Pennsylvania. It was there that Melva graduated high school, and later became the wife of Henry Clauss Ettinger.
Before she married however, in 1920 when she was only 17 her mother died. In between high school graduation and marriage, she pursued one of the few occupations then open to women, moving for some years to live with her
Aunt Dell
in Philadelphia, where she worked in millinery, making ladies' hats. Aunt Dell, her mother's sister, was probably warm loving support to her and a motherly figure until Melva married. Henry Ettinger was a good prospect, from a family with an established stove, plumbing and heating business, but he also came along at a good time, as Melva was ready to get out of the house when her father was readying to take a new wife which would happen in June 1924. Melva and Henry were married June 7, 1923 by the Rev. John A. Smith in Allentown. Prior to marriage, Melva's address was 43 N. 13th Street in Allentown, and she had turned 20 less than 2 months before her nuptuals.
Once married, Melva became the mother of Ruth Emma (Ettinger) Duncan, Lucille May (Ettinger) Romberger, Edwin H Ettinger, and Jane Louise (Ettinger) Hunt. Initially the young family lived at 1137 Walnut Street in Allentown (a two-story row home), until Melva's husband's favorite uncle, Ed Ettinger, sold them his large three-story home at 14th and Walnut for a very kind price.
Melva was an excellent cook, and kept her growing family well-fed. They were also well-clothed in part due to her skill with needles, and the clothing, as well as knitted and crocheted items she produced. There's a surviving picture of my mother in a striking white gown with opera-length black gloves, dressed to go a ball, and her mom Melva made the dress. We still have a few of her afghans and aprons too!
While she'd had houses most of her married life, she and my grandpa downsized as they grew older, and this apartment they had was wonderful. Located at 15th and Hamilton in Allentown, it overlooked the main drag going into town, and was in a former private home that had been divided up into three apartments. You parked on the street, right next to a leftover "horsehead on a stick"- with a ring -an old tie-up from back when people really drove horses everywhere. There was a wrap-around front porch, and a large fake owl hanging on one side, probably to deter pigeons from roosting. (I always called it "Oggie" after
's companion.)
My grandparents had the whole second floor- a living room, two bedrooms, bathroom, kitchen, dinette and a large back porch with gray-painted boards overlooking a sizeable green backyard. The dinette had a suspended lamp you could pull down from the ceiling and raise when needed. On one windowsill was a small milk-glass hen on her nest, and whenever I visited, I would pick the top up to find a few M & M's inside for me, as though she had laid eggs. There was a small cold room off the kitchen where you could store things, and aside of the living room was an enchanting small circular room surrounded by curved windows. It faced south and east so it was nearly always sunny and warm, and was a quiet place to sit away from everything. There were two chairs, a marble table, and a few small interesting pieces there- matching brass urns from India, a china shoe with angels on it, an old lamp with a pull-chain to turn it on. The marble side table always had a portrait of Aunt Dell on it, right up until Melva would leave this home at the end years of her life.
In her later years, she was my babysitter when my parents went out for the evening, and I would stay in her apartment which was loaded with antiques, lamps, and a charming melodian which she would play on very rare occasions due to its age and delicacy. I don't know if the furniture was silk, but she had an especially lovely ivory sofa, and a carved loveseat with gold cushions where my child portrait was once painted. Next to that was a glass curio filled with old, small collectibles. In spite of all the nice furnishings, and her intrinsic worrying nature, I don't recall her being too picky about the chance of spills, so we snacked there in front of evening TV. Around Lawrence Welk time, a trip would be made to the kitchen. She'd return with a highball for my grandpa, a Schaefer's beer for herself, and an ice cream soda for me, along with one of those little red and white boxes with a cellophane front of skinny Reisman pretzel sticks. A lover of cheese, she often brought in some cheese for herself, sometimes with ring bologna.
Her apartment was the place to be for Halloween and Saint Patrick's Day, because the parades would go right by the front. If you got cold outside on the curb, you went upstairs to warm up, or could just stay up there and see everything from the front window.
We were at her apartment when the end of the Vietnam War was announced. She told me about how at the end of "the war" (presumably WW I or II) people crowded into the streets and all the church bells were ringing. I waited for the same thing to happen, even opened the windows to hear the bells, and heard and saw nothing.
While her health was not the best and her walking was limited by pain, I can recall her taking me on short walks to nearby West Park where we would feed the squirrels and pigeons. She told me about her family, her rush to get out of the house and marry after her mom died and her father remarried, and her father's tragic death from a fall down the stairs. In even later years, our special relationship came to an end as her circulatory problems worsened, vein-swapping operations were done, and finally one of her legs was amputated just below the knee. I watched her grow away from me and become more self-focused. As an adult I understand it, but back then it felt like she no longer had any thoughts outside herself. It must have been very hard for her, and she never really did get her old spirit back. Her apartment then became a place full of gifts and pick-me-ups sent by thoughtful friends and family- cheeses, petit fours, candy, perfume, lotions, bathrobes and nighties.
Her weak attempts at learning to walk again were thwarted. While at Good Shepherd for rehabilitation, she was told to "try the parallel bars" by a staff member who then left the room. After waiting, my grandma concluded she was to do so alone, and tried, whereupon she fell, splitting her leg stump open and necessitating a second amputation, this time above the knee. With the loss of the joint, and the artificial leg technology of the time, coupled with her loss of will, she never walked again. Her health worsened to where the help she had from hired folks and my mom were not enough and she needed more care, so she moved to Liberty Nursing Home. I do not recall how long she was there until she had a fall, and in x-raying her after the fall for rib pain, a mass was discovered, and she later died from cancer.
Right before her end, her mind went back in time, as many such people's minds do, to a more pleasant place and time. She spoke of going onto the back porch and having lemonade. I hope somehow she was able to stay there in her mind, so her passing might have felt filled with some comfort.
Family links:
William H Helfrich (1867 - 1939)
Cora Ellen
Helfrich (1874 - 1920)
Henry Clauss Ettinger (1900 - 1971)
Ruth Emma
Duncan (1924 - 1995)
Lucille May
Romberger (1926 - 2012)
Edwin Henry Ettinger (1931 - 1999)
Grace Myrtle
Peters (1900 - 1989)
William Ellsworth Helfrich (1901 - 1964)
Harold Henry Helfrich (1902 - 1982)
Melva Ellen
Ettinger (1903 - 1977)
Claud Morton Helfrich (1911 - 1912)
Calculated relationship
Greenwood Cemetery
Lehigh County
Pennsylvania, USA
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Record added: Jun 21, 2006
Find A Grave Memorial# 14665130
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Shopping at Santa Monica's Museum of Art
We haven't come across a museum shop that we'd like to plunder for a while, but now Santa Monica's Museum of Art Shop does it for us. Quirky, amusing, hip.
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We know just the right recipients for this Cats Attack scratching post: "Does Fluffy have dreams of grandeur? Our new cardboard cityscape scratcher can help her fulfill those dreams by allowing her to reenact her favorite 1950s horror film. Cat Attack, designed by Mat Lyon for Luckies of London is a natural scratch post designed to keep kitty's destructive nails from your upholstery."
A little late for Father's Day, but perhaps a birthday. Gent's Deck: 50 Style & Etiquette tips, in your pocket. "Perfect for those situations in life where you need an ace up your sleeve. From proper revolving door etiquette to learning how to tie a Windsor Knot The Gentleman's Deck will help you become a man of the 21st century."
In this age of video games, apps and overly expensive toys, blocks still win. Here are four sets of blocks to play with: Hebrew, Chinese, Korean and Japanese. "This Hebrew aleph-bet block set offers a great way to educate a child with family heritage as well as learning a language. This set includes 27 blocks made from sustainable Michigan basswood, featuring Hebrew letters, numerals, and animal pictures."
Korean Blocks Lil' Genius: "Learn a new language, or perfect the one you know with this stunning set, made from sustainable Michigan basswood. The Korean character 32 block set includes 22 animals, the numbers 1-10 in English and Korean, a word construction grid, all of the Korean consonants and vowels, and a puzzle. 32 1.75" cubes (44mm)block set includes 22 animals, the numbers 1-10 in English and Korean, a word construction grid, all of the Korean consonants and vowels, and a puzzle. 32 1.75" cubes (44mm) Ages 2+."
Pages: 1 · 2
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|Oct. 1
|Visteon participates in University of Michigan-Dearborn 2015 Fall Career Fair,
Dearborn, MI
|Oct. 8
|Chris Andrews, emerging technologies leader, is a panel speaker on "Cooperative ITS Deployment Challenges" at ITS World Congress, Bordeaux, France
|Oct. 8
|Striding to End Cancer 5K walk/run to support Karmanos Cancer Institute's Jessie Bernard Patient Assistance Fund, Van Buren Twp., MI
|Oct. 20-22
|Tim Yerdon, design, marketing and connected services, is panel speaker on "Marketing to Tech-Centric Buyers" at JD Power Automotive Marketing Roundtable, Las Vegas, NV
3-D Hits Your Dashboard
Sit in your car and compare your 65-inch 4K Ultra HD smart TV that supports 3-D with your vehicle display. Even if your car uses colorful digital graphics, most displays show a flat, two-dimensional image. Take heart. The auto industry is working hard to adopt the trends erupting in consumer electronics.
Visteon Multi-Layer Display Cluster Concept
This concept enables distinctive, visually striking 3-D graphics using two displays in one instrument cluster – a traditional thin-film transistor (TFT) in the back and transparent display in the front. The physical separation of the displays enables the user to see images on two distinct planes, providing a sense of depth which allows for more effective visualization of important driver information elements.


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Young brunette pumped with cock before she eats a nice load of nut butter
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For Immediate Release: 1/30/2014
|Two Millionth Call Milestone for Montgomery County's Online and Telephone Customer Service System
Montgomery County Executive Isiah Leggett today visited the County's MC311 Customer Service Center to congratulate Customer Service Representative Tamara Tyler who, on January 24, fielded a call from the two-millionth customer. Stuart Helfman, a Gaithersburg resident, who had called earlier in the day to report a dead deer behind his house, called back in the afternoon to check on the progress of his report. He was advised by Tyler that Animal Services would handle his request within 24 hours. Helfman's afternoon call was the two-millionth call to the call center.
To celebrate this major milestone, Leggett presented both the caller and the call taker with a one-year pass to the County's indoor aquatic centers.
The MC311 system was launched on June 17, 2010 implementing the centralized Customer Service Center to enhance public access to community services with one easy-to-remember number and a website available 24/7.
"MC311 puts all of Montgomery County's services and information at our residents' fingertips whether online or on the telephone," said Leggett. "Over the past several years, call volume has continued to rise as does the website's online views. We have provided a valuable service to our residents and I am happy that customer satisfaction has been so high."
Since the customer service center opened approximately three-and-a-half years ago, call volume has averaged more than 40,000 calls per month. The 311 web portal, www.MC311.com, which allows residents to find answers and check on the status of service requests, has consistently averaged 25,000 views per month. The most recent Customer Satisfaction Survey of hundreds of MC311 call center users showed that more than 85 percent of callers were satisfied with their overall 311 experience.
Features of the MC311 Customer Service Center include:
• Customers can call one easy-to-remember number or the 10-digit number, 240-777-0311, to access Montgomery County information and services.
• Customers can access information and create or track service requests created on the website at www.MC311.com.
• Weekday operations from 7 a.m. to 7 p.m. The call center is closed on Saturdays, Sundays and holidays.
• Trained Spanish-speaking representatives plus a language interpretation service for other languages.
• TTY access by calling 301-251-4850.
• A state-of-the-art database of information and services about County government that can be continually updated.
• A system that quickly incorporates real time information in case of a weather-related incident or community event/emergency, like a water main break, H1N1 flu epidemic.
For more information about MC311, call the Office of Public Information at 240-777-6507 or visit www.MC311.com.
|Release ID: 14-041
Media Contact: Sue Tucker 240-777-6507
Return to News Releases


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$3,009 zsz 2007 suzuki gsx-r 1000 zsdSuzuki · Tiverton, RIEngine type Four-stroke, liquid-cooled, DOHC, in-line four Bore x stroke 73.4 mm x 59 mm Engine displacement 999 cm Compression ratio 12....Juneds J. · 46 minutes ago
$3,009 zsz 2007 suzuki gsx-r 1000 zsdSuzuki · Rapid City, SDin favor of 1000cc fours. But those 150-horsepower literbikes can be too much on the street, and peaky, 15,000-rpm middleweights aren't a...Jeffreyds J. · 1 hour ago
$3,009 zsz 2007 suzuki gsx-r 1000 zsdSuzuki · Ooltewah, TNarrower fairing and more compact, 4.5-gallon fuel tank are identical to the smaller GSX-R's. The 750's tach wears a 14,000-rpm redline th...Kathrynds K. · 1 hour ago
$18,999 2012 BMW K1600 GTL Premium M432BMW · Marlette, MIFor Sale 2012 BMW K1600 GTL Premium, this bike is hardly broken in with just 7,054 miles. BMW has the process of engineering great tourin...Bennett Powersports · 1 hour ago
$3,009 zsz 2007 suzuki gsx-r 1000 zsdSuzuki · Round Rock, TXtorque and throttle response in the low-to-mid RPM range. The Suzuki Drive Mode Selector (S-DMS) offers push-button selection of two raci...Carolineds C. · 1 hour ago
$3,009 zsz 2007 suzuki gsx-r 1000 zsdSuzuki · West Jordan, UTis a race-developed, lightweight Showa Big Piston front-Fork (BPF) and Single Showa rear shock that features externally adjustable reboun...Joeyds J. · 1 hour ago
$4,500 2014 Honda Shadow Phantom - One OwnerHonda · Ann Arbor, MIHere is a clean 2014 Honda Shadow Phantom 750. Murdered out black motorcycle with only one adult owner and 6957 total miles. Leather sadd...John C. · 6 hours ago
$3,009 zsz 2007 suzuki gsx-r 1000 zsdSuzuki · Burlington, VTBore Stroke: 70.0 mm (2.756 in) x 48.7 mm (1.917 in) Compression Ratio: 12.5 : 1 Engine: 750cc (45.8 cu. In), 4-stroke, liquid-cooled, DO...Warrends W. · 2 hours ago
$3,000 1947 Harley Davidson 45 WLHarley-Davidson · Rocklin, CAHarley Davidson 45 WL with beautiful red and black paint.No expenses spared and no shortcuts taken. Starts easily and drives very well. S...Willieds W. · 2 hours ago
$3,009 zsz 2007 suzuki gsx-r 1000 zsdSuzuki · Skokomish Nation, WABore × stroke: 2.93 inch × 2.26 inch (oversquare - shortstroke) Cooling system: Water cooled Throttle: Cable operated Valves Valve train...Lanceds L. · 2 hours ago
$3,009 zsz 2007 suzuki gsx-r 1000 zsdSuzuki · Hurricane, WVBuilt around the same inline-four engine that was found in the 2008 Suzuki GSX-R1000, the GSX-S1000 has been tuned for street use, though...Tammyds T. ·1 listing · 3 hours ago
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Good going Alan, sounds like you got a good guy there! I look forward to seeing you get a result soon!
Dreaded Fuel Pump Failure On 2003 1.8 Tddi
Started by dougy giro, Nov 05 2013 02:36 PMfuel pump ECU
15 replies to this topic
Posted 13 November 2013 - 03:30 PM
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